cyclic-gmp and Pulmonary-Fibrosis

Pulmonary fibrosis is a progressive and lethal lung disease characterized by accumulation of extracellular matrix and loss of pulmonary function. No cure exists for this pathologic condition, and current treatments often fail to slow its progression or relieve its symptoms. Relaxin was previously shown to induce a matrix-degrading phenotype in human lung fibroblasts in vitro and to inhibit pulmonary fibrosis in vivo. A novel peptide that targets the relaxin RXFP1/LGR7 receptor was recently identified using our computational platform designed to predict novel G protein-coupled receptor peptide agonists. In this study, we examined the antifibrotic properties of this novel peptide, designated CGEN25009, in human cell-based assays and in a murine model of bleomycin-induced pulmonary fibrosis. Similar to relaxin, CGEN25009 was found to have an inhibitory effect on transforming growth factor-β1-induced collagen deposition in human dermal fibroblasts and to enhance MMP-2 expression. The peptide's biological activity was also similar to relaxin in generating cellular stimulation of cAMP, cGMP, and NO in the THP-1 human cell line. In vivo, 2-week administration of CGEN25009 in a preventive or therapeutic mode (i.e., concurrently with or 7 days after bleomycin treatment, respectively) caused a significant reduction in lung inflammation and injury and ameliorated adverse airway remodeling and peribronchial fibrosis. The results of this study indicate that CGEN25009 displays antifibrotic and anti-inflammatory properties and may offer a new therapeutic option for the treatment of pulmonary fibrosis.

Topics: Animals; Bleomycin; Bronchi; Cell Line, Tumor; Collagen; Cyclic AMP; Cyclic GMP; Fibroblasts; Goblet Cells; Humans; Lung; Male; Matrix Metalloproteinase 2; Mice; Mice, Inbred C57BL; Monocytes; Muscle, Smooth; Nitric Oxide; Peptides; Peroxidase; Pulmonary Fibrosis; Receptors, G-Protein-Coupled; Receptors, Peptide; Relaxin; Signal Transduction; Thiobarbituric Acid Reactive Substances; Transforming Growth Factor beta1; Tyrosine

Idiopathic pulmonary fibrosis is an incurable fibrosing disorder that progresses relentlessly to respiratory failure. We hypothesized that a product of heme oxygenase activity, carbon monoxide (CO), may have anti-fibrotic effects. To test this hypothesis, mice treated with intratracheal bleomycin were exposed to low-concentration inhaled CO or ambient air. Lungs of mice treated with CO had significantly lower hydroxyproline accumulation than controls. Fibroblast proliferation, thought to play a central role in the progression of fibrosis, was suppressed by in vitro exposure to CO. CO caused increased cellular levels of p21(Cip1) and decreased levels of cyclins A and D. This effect was independent of the observed suppression of MAPK's phosphorylation by CO but was dependent on increased cGMP levels. Further, CO-exposed cells elaborated significantly less fibronectin and collagen-1 than control cells. This same effect was seen in vivo. Suppression of collagen-1 production did not depend on MAPK or guanylate cyclase signaling pathways but did depend on the transcriptional regulator Id1. Taken together, these data suggest that CO exerts an anti-fibrotic effect in the lung, and this effect may be due to suppression of fibroblast proliferation and/or suppression of matrix deposition by fibroblasts.

Topics: Animals; Bleomycin; Carbon Monoxide; Cell Cycle; Cell Division; Cyclic AMP; Cyclic GMP; Hydroxyproline; Male; Mice; Mice, Inbred CBA; Pulmonary Fibrosis

The purpose of this study was to evaluate the pathophysiologic role of atrial natriuretic peptide (ANP) as a pulmonary artery vasodilator in patients with acute respiratory failure receiving artificial ventilation. Twenty-one consecutive patients were studied, 12 without and 9 with preexisting cardiopulmonary disease. Pulmonary artery plasma ANP levels were significantly higher than the levels obtained in the superior vena cava and radial artery. Plasma ANP levels correlated significantly with the plasma levels of its second messenger, guanosine 3',5'-cyclic monophosphate (cGMP). In the 12 patients without prior cardiopulmonary disease, plasma ANP levels correlated significantly with mean pulmonary arterial pressure (MPAP). This correlation was not found in the nine patients with preexisting cardiopulmonary disease. The cGMP/ANP ratio, indicating the biologic effect of ANP, was also higher in the patients without preexisting cardiopulmonary disease. These results are compatible with clearance and vasodilator activity of ANP in the pulmonary vascular bed, but only in patients without preexisting cardiopulmonary disease.

Topics: Acute Disease; Adult; Aged; Aged, 80 and over; Atrial Function, Right; Atrial Natriuretic Factor; Blood Pressure; Cyclic GMP; Female; Heart Failure; Humans; Lung Diseases, Obstructive; Male; Middle Aged; Pulmonary Artery; Pulmonary Fibrosis; Radial Artery; Respiration, Artificial; Respiratory Insufficiency; Vena Cava, Superior

Cyclic nucleotides play an important role in the regulation of fibroblast proliferation and collagen metabolism. In the present study, the antifibrotic potential of dibutyrylcAMP (Bt2cAMP) was evaluated in the bleomycin (BLM)-hamster model of pulmonary fibrosis. Bt2cAMP (10 mg kg-1, s.c.) or saline (SA, s.c.) was given daily two days prior to the first intratracheal (i.t.) dose of BLM or SA and thereafter throughout the study. BLM or SA was instilled i.t. in three consecutive doses (2.5, 2.0 and 1.5 U 5ml-1 kg-1) at weekly intervals. Hamsters were killed at 7, 14 and 20 days after the third i.t. instillation. Bt2cAMP significantly reduced the contents of lung hydroxyproline and lung thiobarbituric acid reactive substance equivalents in BLM-treated animals at 7 and 14 days. Bt2cAMP significantly elevated lung superoxide dismutase activity in BLM-treated animals at 7 days. Lung prolyl hydroxylase activity was significantly elevated at 14 and 20 days in SABLM- and Bt2cAMPBLM-treated animals. The ratio of cAMP/cGMP was significantly reduced at all time points in SABLM-treated animals but only at 7 and 14 days in Bt2cAMPBLM-treated animals. Bt2cAMP caused no significant changes in lung calcium and calmodulin levels and protein content of the bronchoalveolar lavage. BLM significantly increased various inflammatory cell counts in the lavage at all three time points. The cell counts in the Bt2cAMPBLM groups were generally lower at 7 days and higher at 20 days than those of the SABLM groups. Histological evaluation showed that the lungs of Bt2cAMPBLM-treated hamsters progressed from an inflammatory cell lesion to a fibrotic lesion at a slower rate than the SABLM groups. It was concluded that Bt2cAMP attenuated BLM-induced pulmonary fibrosis in hamsters in part by delaying the acute phase of the inflammatory reaction.

Topics: Animals; Bleomycin; Bronchoalveolar Lavage Fluid; Bucladesine; Cricetinae; Cyclic AMP; Cyclic GMP; Hydroxyproline; Lung; Male; Mesocricetus; Pulmonary Fibrosis; Superoxide Dismutase

Cyclic nucleotides have been shown in vitro to regulate fibroblast proliferation and/or collagen production. We have reported previously that propranolol, which decreases the cAMP/cGMP ratio, potentiates the amount of fibrosis produced in a damaged lung. The purpose of this study was to determine if elevations in the cAMP/cGMP ratio may attenuate collagen production by fibroblasts following lung damage. Lung injury was induced in mice by either butylated hydroxytoluene (BHT) (350 or 400 mg/kg intraperitoneally) or bleomycin (4 units/kg intratracheally). The mice were treated with a phosphodiesterase inhibitor, aminophylline (20 mg/kg twice daily), prior to induction of lung injury and for the duration of the study. Cyclic nucleotide changes in the lung were also determined during lung injury, with and without aminophylline. The administration of aminophylline, which increased the cAMP/cGMP ratio, resulted in attenuation of the increase in total lung collagen normally seen after injury, while having no effect on collagen levels in the undamaged lung. The results are compatible with the hypothesis that elevation of whole lung cAMP/cGMP ratio early in the damage and repair process correlates with decreased hydroxyproline deposition.

Topics: Aminophylline; Animals; Bleomycin; Butylated Hydroxytoluene; Collagen; Cyclic AMP; Cyclic GMP; Hydroxyproline; Lung; Male; Mice; Pulmonary Fibrosis; Time Factors

Topics: Animals; Cyclic GMP; Guanylate Cyclase; Guinea Pigs; Lung; Male; Paraquat; Pulmonary Fibrosis